Production process of rhamnolipid daily chemical raw material

Abstract

Provided is a production process of a rhamnolipid daily chemical raw material. By controlling the acidification pH of a rhamnolipid fermentation broth, three solids, i.e., solid A, solid B and solid C each having a different ratio of mono-rhamnolipid to di-rhamnolipid are obtained; the solid A, the solid B and the solid C are separately dissolved by using an alcohol solvent, and filtered; the solvent in the filtrate is evaporated to dryness to correspondingly obtain a solid a, a solid b and a solid c; and then the solid a, the solid b and the solid c are compounded with a KOH solution and 1,3-butanediol to obtain three surfactant product solutions, i.e., solution a, solution b and solution c each having different pH and CMC, which can be applied to different application fields.

Description

Production process of rhamnolipid daily chemical raw material

[0001] Cross-reference to related applications

[0002] This application claims priority to the Chinese patent application No. 202410873229.X, filed on July 1, 2024, entitled “Production process of rhamnolipid daily chemical raw material”, the entire content of which is incorporated herein by reference. TECHNICAL FIELD

[0003] The present application belongs to the field of fermentation engineering, and relates to a production process of rhamnolipid daily chemical raw material. BACKGROUND

[0004] Rhamnolipid can be metabolized by Pseudomonas aeruginosa under certain environmental conditions. It has good surface activity and interfacial activity, and can be widely used in petrochemical, environmental, pharmaceutical, food and agricultural fields. It is a non-toxic and biodegradable biological surfactant with the longest research time, the best effect and the most mature application technology. Rhamnolipid is divided into four types according to its structure: disaccharide double lipid, disaccharide single lipid, monosaccharide double lipid and monosaccharide single lipid. Disaccharide double lipid and disaccharide single lipid are combined as disaccharide lipid, and monosaccharide double lipid and monosaccharide single lipid are combined as monosaccharide lipid. Disaccharide lipid has lower surface tension, better foam performance and lower pH application range than monosaccharide lipid, and is more suitable for application in personal care field. In the current production process of rhamnolipid, the ratio of monosaccharide lipid to disaccharide lipid of the obtained rhamnolipid is fixed. The ratio of disaccharide lipid of the product of Winclean is about 95%, and the ratio of disaccharide lipid of the products of domestic manufacturers such as Deguo Bio and Xin Yinxiang is about 50-60%. The ratio of monosaccharide lipid to disaccharide lipid cannot be regulated according to the downstream demand to meet the needs of customers in different fields. Once the ratio of monosaccharide lipid to disaccharide lipid after fermentation is determined, the ratio of monosaccharide lipid to disaccharide lipid of the final product is also determined, and different products cannot be flexibly switched to meet the needs of different application fields.

[0005] SUMMARY

[0006] The technical problem to be solved by the present application is that in the prior art, different rhamnolipid products with different ratios of disaccharide lipid cannot be obtained by one separation process. By controlling different pH during the acidification crystallization process of rhamnolipid, products with different ratios of disaccharide lipid are obtained.

[0007] The inventors of the present application found that the solubility of rhamnolipids decreases under acidic conditions and the rhamnolipids are converted into crystals, the sensitivity of di-rhamnolipids to pH is higher than that of mono-rhamnolipids, under certain pH conditions, di-rhamnolipids have been crystallized, but mono-rhamnolipids still have good solubility in the solution, by this feature, the pH gradient of the solution can be controlled to control the proportion of di-rhamnolipids in the product, so that a process route for preparing rhamnolipid products of different specifications using a specification of rhamnolipid fermentation broth is realized.

[0008] Further, the rhamnolipids finally exist in the form of anionic surfactants in the final product, in order to maintain good surface activity and solubility, a certain proportion of 1,3-butanediol and KOH is added in the final product, the resulting formula product has better compatibility with other daily chemical raw materials, and no preservative needs to be added.

[0009] In a first aspect, the present application provides a production process of rhamnolipid daily chemical raw material, comprising the following steps:

[0010] (1) Pseudomonas aeruginosa is fermented and cultured, the pH in the fermentation and culture process is controlled at 7.0-8.0, a fermentation broth with a rhamnolipid content of 60-80 g / L and a di-rhamnolipid proportion of 60-70% in the rhamnolipids is obtained;

[0011] (2) a solid-liquid separation device is used to remove the bacteria in the fermentation broth, to obtain a sterilized fermentation broth;

[0012] (3) acid is added to the sterilized fermentation broth to adjust the pH to 3.0-3.5, and solids are precipitated, the solid-liquid phase is separated, to obtain solids A and supernatant A; then acid is continuously added to the supernatant A to reduce the pH by 0.5, and solids are precipitated, the solid-liquid phase is separated, to obtain solids B and supernatant B; then acid is continuously added to the supernatant B to reduce the pH by 0.5 again, and solids are precipitated, the solid-liquid phase is separated, to obtain solids C and supernatant C.

[0013] In some embodiments, in the above production process, the mass percentage of rhamnolipids in the solids A is 80-93%, and the mass percentage of di-rhamnolipids in the rhamnolipids is 90-95%;

[0014] the mass percentage of rhamnolipids in the solids B is 80-93%, and the mass percentage of di-rhamnolipids in the rhamnolipids is 70-80%; and / or

[0015] the mass percentage of rhamnolipids in the solids C is 80-93%, and the mass percentage of di-rhamnolipids in the rhamnolipids is 35-50%.

[0016] In some embodiments, in any of the production processes described above, the yield of solid A is 18-20%, the yield of solid B is 28-30%, the yield of solid C is 43-45%, and the overall yield is 89-95%.

[0017] In some embodiments, in any of the production processes described above, in step (1), the fermentation culture uses soybean oil as the carbon source and yeast powder as the nitrogen source, the fermentation temperature is 30-37°C, the aeration volume is 0.9-2.5vvm, the stirring speed is 150-500rpm, and the fermentation time is 168-196 hours.

[0018] In some embodiments, in any of the production processes described above, in step (1), the fermentation medium used in the fermentation culture is: soybean oil 15-25g / L, yeast powder 10-17g / L, dipotassium phosphate 2-3.5g / L, disodium hydrogen phosphate 5-6.2g / L, magnesium sulfate 0.1-0.25g / L, sodium chloride 1-2.5g / L, pH 7.0-7.5, and the balance is water.

[0019] In some embodiments, in any of the production processes described above, in step (1), a feed medium is added during the period from the 48th hour of fermentation to 24 hours before the end of fermentation, so that the concentration of soybean oil is not less than 10g / L. In some embodiments, the feed medium is: soybean oil 10-15g / L, yeast powder 5-8g / L, dipotassium phosphate 0.2-0.5g / L, disodium hydrogen phosphate 0.1-0.2g / L, magnesium sulfate 0.05-0.1g / L, sodium chloride 1-1.5g / L, pH 7.0-7.5, and the balance is water.

[0020] In some embodiments, in any of the production processes described above, before the fermentation culture, there is a step of seed culture, in which the P. aeruginosa is inoculated into a seed culture medium and cultured at 35-37°C and 150-200rpm until the OD600 reaches about 4 to obtain a seed liquid; the seed liquid is inoculated into the fermentation medium at an inoculation amount of 3-5% (volume ratio). In some embodiments, the seed culture medium is: yeast powder 5-8g / L, peptone 10-12g / L, sodium chloride 0.5-1g / L, and the balance is water.

[0021] In some embodiments, in any of the production processes described above, in step (2), the solid-liquid separation device can be a positive pressure filter, a centrifuge, a plate and frame filter, a ceramic membrane, or an organic membrane.

[0022] In some embodiments, in any of the production processes described above, in step (3), the acid is an organic acid and / or an inorganic acid.

[0023] Optionally, the organic acid is one or more of citric acid, formic acid, acetic acid, valeric acid, butyric acid, optionally, the organic acid is acetic acid;

[0024] Optionally, the inorganic acid is one or more of sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, optionally, the inorganic acid is sulfuric acid;

[0025] The solid-liquid phase can be separated by centrifugation.

[0026] In some embodiments, the production process described above further comprises:

[0027] (4) dissolving the solid A, solid B and solid C respectively using an alcohol solvent, filtering to obtain a filtrate, evaporating the solvent in the filtrate to obtain a solid a, a solid b and a solid c.

[0028] In some embodiments, in the production process described above, in step (4), the alcohol solvent is one or more of methanol, ethanol, propanol and butanol;

[0029] The alcohol solvent is used in an amount of 1.5-3 times the mass of the solid to be dissolved;

[0030] The temperature of the dissolution is 20-40℃, and the time is 1-3 hours; and / or

[0031] The filtering uses plate and frame filtration, the filter cloth has a pore size of 1-5 μm, and the material can be selected from polypropylene, polyether sulfone and nylon.

[0032] In some embodiments, the production process described above further comprises:

[0033] (5) adding to the solid a an aqueous KOH solution and / or an aqueous NaOH solution with a mass percentage of 5-6% and 1,3-butanediol, glycerol or 1,2-pentanediol with a mass of 0.5-0.55 times that of the solid a, to obtain a solution a with a pH of 5.5-6.0, the mass percentage of rhamnolipids in the solution a being 35-40%, and the mass percentage of disaccharolipids in the rhamnolipids being 90-95%;

[0034] adding to the solid b an aqueous KOH solution and / or an aqueous NaOH solution with a mass percentage of 5-6% and 1,3-butanediol, glycerol or 1,2-pentanediol with a mass of 0.6-0.65 times that of the solid b, to obtain a solution b with a pH of 6.0-6.5, the mass percentage of rhamnolipids in the solution b being 32-35%, and the mass percentage of disaccharolipids in the rhamnolipids being 70-80%; and / or

[0035] The solid c is added with 1.3-1.4 times of its mass of a 5-6% mass percentage KOH aqueous solution and / or a 5-6% mass percentage NaOH aqueous solution and 0.7-0.75 times of its mass of 1,3-butanediol, glycerol or 1,2-pentanediol to obtain a solution c with a pH of 6.5-7.0, a mass percentage of rhamnolipid of 30-33% and a mass percentage of di-rhamnolipid in the rhamnolipid of 35-50%.

[0036] In some embodiments, in the above production process, the pH of the solution a is resistant to 5-6 and the CMC is 0.005-0.007 mol / L; it is suitable for application in skin care products (such as facial cleanser) in the field of personal care;

[0037] The pH of the solution b is resistant to 6.5-7.5 and the CMC is 0.008-0.01 mol / L; it is suitable for application in hair care products (such as shampoo) in the field of personal care; and / or

[0038] The pH of the solution c is resistant to 7.5-8.5 and the CMC is 0.012-0.02 mol / L; it is suitable for application in laundry liquid in the field of home care.

[0039] In the second aspect, the present application provides the solid A, solid B, solid C, solid a, solid b, solid c, solution a, solution b and solution c obtained by any of the above production processes.

[0040] In the third aspect, the present application provides the use of the obtained solid A, solid a and / or solution a as a daily chemical raw material for preparing skin care products (such as facial cleanser) in the field of personal care.

[0041] In the fourth aspect, the present application provides the use of the obtained solid B, solid b and / or solution b as a daily chemical raw material for preparing hair care products (such as shampoo) in the field of personal care.

[0042] In the fifth aspect, the present application provides the use of the obtained solid C, solid c and / or solution c as a daily chemical raw material for preparing laundry liquid in the field of home care.

[0043] In some embodiments, in any of the above uses, the daily chemical raw material is added in an amount of 2-5 wt% in the personal care product or the home care product, calculated based on the rhamnolipid contained therein.

[0044] The technical solution of the present application has the following advantages:

[0045] (1) The application provides a process for separating different mono and double glycolipid products by controlling the acidification pH of rhamnolipid fermentation liquor, which can obtain various rhamnolipid products with different mono and double glycolipid ratios without changing the fermentation conditions.

[0046] (2) The application also provides rhamnolipid complex products, which are complexed with, for example, KOH and 1,3-butanediol to increase the solubility of rhamnolipid in the system, obtain products with different pH and CMC, and have good preservative effect, without the need for additional addition of preservatives in application, which can be applied in different fields. DETAILED DESCRIPTION

[0047] The application will be further described below in conjunction with specific examples. It should be understood that the following examples are only used to illustrate the application and are not used to limit the scope of the application. Unless otherwise specified, the technical means used in the examples are conventional operations in the art, or according to the experimental methods suggested by the manufacturers of the kits and instrument equipment. The reagents and biological materials used in the examples, unless otherwise specified, can be obtained commercially.

[0048] In the application, the fermentation equipment used is: Iversen 7.5L fermenter;

[0049] The reagents used and sources are as follows:

[0050] Soybean oil (premium grade), purchased from COFCO Corporation;

[0051] Concentrated sulfuric acid, 98% content, analytical pure, purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd.;

[0052] 1,3-Butanediol, analytical pure, purchased from Aldrin;

[0053] Lauryl glucoside, cocamidopropyl betaine, glycerol, sodium laureth sulfate, cocodiethanolamide, sodium fatty alcohol ether sulfate, sodium dodecyl sulfate, disodium ethylenediaminetetraacetate, sodium carbonate, sodium chloride, sodium benzoate, all purchased from Beijing Inokai Technology Co., Ltd.;

[0054] Rose essential oil, essence, all purchased from AIP Perfume Group Co., Ltd.;

[0055] Grape seed extract, purchased from Xi'an Zebang Biological Technology Co., Ltd.;

[0056] Pearlescent agent, cationic guar gum, both purchased from Heda Chemicals (Shanghai) Co., Ltd.;

[0057] Silicone emulsion, purchased from Shin-Etsu Silicone (Nantong) Co., Ltd.

[0058] The detection method of rhamnolipid content is sulfuric acid-anthrone method, and the specific operation method is as follows:

[0059] 1) Preparation of anthrone solution: 0.2 g of anthrone was dissolved in 100 mL of 80% sulfuric acid, and stored in the dark for use. The solution should be prepared immediately before use and not stored.

[0060] 2) 0.5 mL of the sample to be tested was placed in a 10-15 mL graduated test tube with a stopper, and cooled in an ice water bath. 2 mL of the anthrone solution was added, and mixed quickly (to minimize the reaction). The mixture was then placed in a boiling water bath for 10 minutes, removed and cooled in an ice water bath to room temperature, and the absorbance was measured at 620 nm.

[0061] 3) Rhamnose standard curve: 0.1 g of rhamnose was weighed into a 250 mL volumetric flask, and diluted to volume with water to prepare a rhamnose stock solution of 400 mg / L. The stock solution was then diluted to obtain different concentrations of the standard samples, as shown in Table 1.

[0062] Table 1

[0063] The absorbance was measured using the anthrone method, and a standard curve was prepared.

[0064] y = ax + b

[0065] where y is the rhamnose content, and x is the absorbance at 620 nm.

[0066] The final rhamnolipid content was calculated using the following formula:

[0067] Rhamnolipid content = rhamnose content in the reaction solution x dilution factor x 3.4.

[0068] The ratio of rhamnolipid monomer to rhamnolipid dimers was determined using high performance liquid chromatography (HPLC), and the area normalization method was used to determine the ratio of different configurations. The HPLC detection conditions were as follows:

[0069] Chromatographic column: Agilent C18 reverse phase chromatographic column, ZORBAX Eclipse XDB-C18 (4.6 x 250 mm / 5 μm);

[0070] Mobile phase: a mixture of methanol, water and formic acid prepared according to the ratio of 600 g of methanol, 250 g of water and 0.5 mL of formic acid;

[0071] Flow rate: 1 mL / min;

[0072] Injection volume: 10 μL;

[0073] Elution: isocratic elution for 40 min;

[0074] Detector: differential detector;

[0075] Column temperature: 35°C.

[0076] For example, solid b in Example 1 was formulated into a solution with a concentration of 1% (1 g / 100 ml) using methanol, and was subjected to the above-mentioned high performance liquid chromatography, and the retention time of rhamnolipid was shown in Table 2.

[0077] Table 2

[0078] Rha-C8C 10 / C 10 C8, Rha-C 10 C 10 and Rha-C 12 C 10 / C 10 C 12 Rhamnolipid of the type Rha-C m C n is shown below; for Rha-C8C 10 , m is 8 and n is 10; for Rha-C 10 C8, m is 10 and n is 8; for Rha-C 10 C 10 , m is 10 and n is 10; for Rha-C 12 C 10 , m is 12 and n is 10; and for Rha-C 10 C 12 , m is 10 and n is 12.

[0079] Rha = C 12 C 10 is shown below; for Rha = C m C n , the bond with a dotted line is a double bond (the double bond can exist between any two methylene groups of C m (in other words, the fatty acid connected to the sugar ring is a monounsaturated fatty acid) in addition to the positions identified), m is 12 and n is 10.

[0080] RhaRha-C8C 10 / C 10 C8, RhaRha-C 10 C 10 and RhaRha-C 12 C 10 / C 10 C 12 Rhamnolipid of the type RhaRha-C m C n is shown below; for RhaRha-C8C 10 , m is 8 and n is 10; for RhaRha-C 10C8, m is 10, n is 8; for RhaRha-C 10 C 10 m is 10, n is 10; for RhaRha-C 12 C 10 m is 12, n is 10; for RhaRha-C 10 C 12 m is 10 and n is 12.

[0081] RhaRha=C 10 C 12 The structural formula is as follows: RhaRha=C m C n As shown, the keys marked with dashed lines are double bonds (these double bonds can exist not only at the marked positions, but also in C). m Between any two methylene groups (i.e., the fatty acid connected to the sugar ring is a monounsaturated fatty acid), m is 10 and n is 12.

[0082] The monoglycolipids in this application refer to Rha-C8C 10 / C 10 C8, Rha-C 10 C 10 Rha=C 12 C 10 and Rha-C 12 C 10 / C 10 C 12 The sum, disaccharides refer to RhaRha-C8C 10 / C 10 C8, RhaRha-C 10 C 10 RhaRha=C 10 C 12 and RhaRha-C 12 C 10 / C 10 C 12 The sum of .

[0083] CMC stands for Critical Micelle Concentration, which measures the critical concentration at which a surfactant can form micelles in a solution. The detection method is the surface tension method, and the instrument used is a surface tension meter.

[0084] All culture media in the following examples were sterilized at 121°C for 20 minutes.

[0085] Unless otherwise specified, the contents, proportions, and percentages in the following examples are all mass percentages.

[0086] Example 1

[0087] (1) Rhamnolipid fermentation process

[0088] Seed medium: yeast powder 8 g / L, peptone 12 g / L, sodium chloride 1 g / L, the rest being water.

[0089] Fermentation medium: soybean oil 25 g / L, yeast powder 17 g / L, dipotassium hydrogen phosphate 3.5 g / L, disodium hydrogen phosphate 6.2 g / L, magnesium sulfate 0.25 g / L, sodium chloride 2.5 g / L, pH adjusted to 7.0 with sodium hydroxide, the rest being water.

[0090] Feed medium: soybean oil 15 g / L, yeast powder 8 g / L, dipotassium hydrogen phosphate 0.5 g / L, disodium hydrogen phosphate 0.2 g / L, magnesium sulfate 0.1 g / L, sodium chloride 1.5 g / L, pH adjusted to 7.0 with sodium hydroxide, the rest being water.

[0091] Pseudomonas aeruginosa (ATCC 10145) was inoculated into the seed medium and subjected to primary activation culture at 37°C and 200 rpm until the OD600 reached about 4, to obtain a seed solution.

[0092] The seed solution was inoculated into the fermentation medium in a fermenter at an inoculation amount of 5% (by volume), the volume of the fermenter was 7.5 L and the liquid loading was 3 L, and the fermentation culture was carried out at 30°C, 150 rpm and 0.9 vvm aeration, while the pH in the fermentation process was controlled at about 7.0 (pH was adjusted by adding 10% sulfuric acid solution), and the fermentation was carried out for 168 hours, during which the feed medium was added from the 48th hour to 24 hours before the end of the fermentation, so that the concentration of soybean oil was not less than 10 g / L, and finally 3.5 L of fermentation broth was obtained, and the rhamnolipid content in the fermentation broth was 60 g / L and the proportion of diacyl lipids in the rhamnolipid was 60% as determined by sampling.

[0093] (2) The bacteria in the fermentation broth were removed by filtration through a positive pressure filter (filter cloth with 200 mesh size) to obtain a sterilized fermentation broth.

[0094] (3) A 50% sulfuric acid solution was added to the sterilized fermentation broth to adjust the pH to 3.0, and solids were precipitated, which were centrifuged at 10°C and 8000 rpm for 20 min to obtain solids A and supernatant A; then a 50% sulfuric acid solution was further added to the supernatant A to adjust the pH to 2.5, and solids were precipitated, which were centrifuged at 10°C and 8000 rpm for 20 min to obtain solids B and supernatant B; then a 50% sulfuric acid solution was further added to the supernatant B to adjust the pH to 2.0, and solids were precipitated, which were centrifuged at 10°C and 8000 rpm for 20 min to obtain solids C and supernatant C.

[0095] The mass of solid A, solid B and solid C is 40.6 g, 63.2 g and 97.1 g respectively.

[0096] It is detected that the rhamnolipid content of solid A is 93%, and the double sugar lipid accounts for 95% in the rhamnolipid; the rhamnolipid content of solid B is 93%, and the double sugar lipid accounts for 80% in the rhamnolipid; the rhamnolipid content of solid C is 93%, and the double sugar lipid accounts for 50% in the rhamnolipid.

[0097] The yield of solid A is 18%, the yield of solid B is 28%, the yield of solid C is 43%, and the overall yield is 89%.

[0098] The yield calculation formula is: solid A / B / C mass*rhamnolipid content / (rhamnolipid content in fermentation broth*fermentation broth volume) *100%.

[0099] (4) The solid A, solid B and solid C are dissolved using ethanol with a mass of 3 times the mass of the solid to be dissolved, the dissolution temperature is 20°C, and the dissolution time is 3 hours. After complete dissolution, filtration is performed to remove residual inorganic salts, plate and frame filtration is used, the filter cloth pore size is 5 μm, and the material is polypropylene. The obtained filtrate is evaporated to dryness by reduced pressure distillation, and solid a, solid b and solid c are obtained correspondingly.

[0100] (5) In solid a, 0.9 times the mass of 5% mass percentage KOH aqueous solution and 0.5 times the mass of 1,3-butanediol are added, stirred uniformly, and a solution a of 90 g with pH 5.5 is obtained. The rhamnolipid content of solution a is 40%, and the double sugar lipid accounts for 95% in the rhamnolipid. Solution a is prepared into a rhamnolipid content of 5% aqueous solution, and the measured pH is 5.0. The foam is rich, and the CMC of the product is 0.005 mol / L.

[0101] In solid b, 1.2 times the mass of 5% mass percentage KOH aqueous solution and 0.6 times the mass of 1,3-butanediol are added, stirred uniformly, and a solution b of 150 g with pH 6.0 is obtained. The rhamnolipid content of solution b is 35%, and the double sugar lipid accounts for 80% in the rhamnolipid. Solution b is prepared into a rhamnolipid content of 5% aqueous solution, and the measured pH is 6.5. The foam is moderate, and the CMC of the product is 0.008 mol / L.

[0102] In the solid c, 1.3 times of the mass of the 5% mass percentage KOH aqueous solution and 0.7 times of the mass of the 1,3-butanediol were added, stirred uniformly to obtain a solution c 240 g with pH 6.5, the rhamnolipid content of the solution c was 33%, and the proportion of the di-rhamnolipid in the rhamnolipid was 50%; the solution c was prepared into a rhamnolipid content of 10% aqueous solution, and the pH was 7.5, the foam was less, and the CMC of the product was 0.012 mol / L.

[0103] Example 2

[0104] (1) Rhamnolipid fermentation process

[0105] Seed medium: 5 g / L of yeast powder, 10 g / L of proteose peptone, 0.5 g / L of sodium chloride, and the balance of water.

[0106] Fermentation medium: 15 g / L of soybean oil, 10 g / L of yeast powder, 2 g / L of dipotassium hydrogen phosphate, 5 g / L of disodium hydrogen phosphate, 0.1 g / L of magnesium sulfate, and 1 g / L of sodium chloride, and the balance of water.

[0107] Supplemental medium: 10 g / L of soybean oil, 5 g / L of yeast powder, 0.2 g / L of dipotassium hydrogen phosphate, 0.1 g / L of disodium hydrogen phosphate, 0.05 g / L of magnesium sulfate, and 1 g / L of sodium chloride, and the balance of water.

[0108] Pseudomonas aeruginosa (ATCC 10145) was inoculated into the seed medium and subjected to primary activation culture at 35°C and 150 rpm, and the seed liquid was obtained when the OD600 reached about 4.

[0109] The seed liquid was inoculated into the fermentation medium in the fermentation tank at a 3% (volume ratio) inoculation amount (the volume of the fermentation tank was 7.5 L, and the liquid loading amount was 2 L), and the fermentation culture was carried out at 37°C, 500 rpm, and 2.5 VVM ventilation, while the pH in the fermentation process was controlled at about 8.0 (the pH was adjusted by adding 10% mass percentage sulfuric acid aqueous solution), and the fermentation was ended after 196 hours, and the supplemental medium was added during the period from the 48th hour of the fermentation to 24 hours before the end of the fermentation, so that the concentration of the soybean oil was not less than 10 g / L, and finally the fermentation liquid 2.25 L was obtained, and the rhamnolipid content in the fermentation liquid was 80 g / L, and the proportion of the di-rhamnolipid in the rhamnolipid was 70%.

[0110] (2) The bacteria in the fermentation liquid were removed by filtering through a positive pressure filter (the filter cloth was 200 mesh in size) to obtain the bacteria-removed fermentation liquid.

[0111] (3) In the sterilized fermentation broth, 40% acetic acid aqueous solution was added to adjust the pH to 3.5, and solid was precipitated. Centrifugation was performed at 10°C and 8000 rpm for 20 min to obtain solid A and supernatant A. Then 40% acetic acid aqueous solution was further added to the supernatant A to adjust the pH to 3.0, and solid was precipitated. Centrifugation was performed at 10°C and 8000 rpm for 20 min to obtain solid B and supernatant B. Then 40% acetic acid aqueous solution was further added to the supernatant B to adjust the pH to 2.5, and solid was precipitated. Centrifugation was performed at 10°C and 8000 rpm for 20 min to obtain solid C and supernatant C.

[0112] The mass of solid A, solid B and solid C was 45 g, 67.5 g and 101 g, respectively.

[0113] It was detected that the rhamnolipid content of solid A was 80%, and the double glycolipid accounted for 90% in the rhamnolipid; the rhamnolipid content of solid B was 80%, and the double glycolipid accounted for 70% in the rhamnolipid; the rhamnolipid content of solid C was 80%, and the double glycolipid accounted for 35% in the rhamnolipid.

[0114] The yield of solid A was 20%, the yield of solid B was 30%, the yield of solid C was 45%, and the overall yield was 95%.

[0115] (4) Solid A, solid B and solid C were dissolved in methanol with a mass of 1.5 times the mass of the solid to be dissolved, respectively, and the dissolution temperature was 40°C. The dissolution time was 1 hour. After complete dissolution, filtration was performed to remove residual inorganic salts. Plate and frame filtration was used, and the filter cloth pore size was 1 μm and the material was polyether sulfone. The obtained filtrate was evaporated to dryness by reduced pressure distillation, and solid a, solid b and solid c were obtained correspondingly.

[0116] (5) In solid a, 5% KOH aqueous solution with a mass of 1.1 times the mass of solid a and 1,3-butanediol with a mass of 0.5 times the mass of solid a were added and stirred uniformly to obtain solution a with a pH of 6.0 and a mass of 90 g. The rhamnolipid content of solution a was 35%, and the double glycolipid accounted for 90% in the rhamnolipid. Solution a was prepared into an aqueous solution with a rhamnolipid content of 5%. It was detected that the pH was 5.2, and the foam was abundant. It was detected that the CMC of the product was 0.007 mol / L.

[0117] In solid b, 5% KOH aqueous solution with a mass of 1.25 times the mass of solid b and 1,3-butanediol with a mass of 0.6 times the mass of solid b were added and stirred uniformly to obtain solution b with a pH of 6.5 and a mass of 150 g. The rhamnolipid content of solution b was 32%, and the double glycolipid accounted for 70% in the rhamnolipid. Solution b was prepared into an aqueous solution with a rhamnolipid content of 5%. It was detected that the pH was 6.7, and the foam was moderate. It was detected that the CMC of the product was 0.01 mol / L.

[0118] In the solid c, 1.4 times of the mass of the mass percentage of 5% KOH aqueous solution and 0.7 times of the mass of 1,3-butanediol were added, stirred uniformly to obtain a solution c 240 g with pH 7.0, the rhamnolipid content of the solution c was 30%, and the double glycolipid accounted for 35% in the rhamnolipid; the solution c was prepared into an aqueous solution with a rhamnolipid content of 10%, and the pH was 7.9, the foam was less, and the CMC of the product was 0.02 mol / L.

[0119] Comparative Example

[0120] (1) The rhamnolipid fermentation process of the present comparative example was the same as step (1) of Example 1.

[0121] (2) The fermentation broth was subjected to plate and frame filtration (filter cloth pore size 5 μm, polypropylene), to obtain a sterilized fermentation broth.

[0122] (3) Concentrated sulfuric acid was added to the sterilized fermentation broth to pH 2.5, centrifuged at 10 ℃ and 8000 rpm for 20 min, and the precipitate was collected. The precipitate was a rhamnolipid crude product, the rhamnolipid content of the crude product was 85%, the double glycolipid accounted for 63% in the rhamnolipid, and the yield of the crude product was 75% (yield calculation formula: rhamnolipid crude product mass*rhamnolipid content / (rhamnolipid content in fermentation broth*fermentation broth volume) *100%).

[0123] The crude product was prepared into an aqueous solution with a rhamnolipid content of 10%, and the CMC of the product was 0.25 mol / L.

[0124] Application Example

[0125] Face wash

[0126] The three different double glycolipid ratio daily chemical raw materials (solutions a / b / c) obtained in Example 1 and the rhamnolipid crude product obtained in the comparative example were formulated into a face wash according to the formulation of Table 3.

[0127] Table 3

[0128] Processing process:

[0129] The above raw materials were added according to the formulation ratio, stirred at 35 ℃ for 20 minutes, then the pH of the solution was adjusted to 6.0 with citric acid, and cooled to room temperature.

[0130] Ten consumers were selected to score the foaming property, foam fineness, post-washing moisturizing property and washing power of the face washes respectively added with the solutions a / b / c of Example 1 and the rhamnolipid crude product of the comparative example, with a full score of 10. The results are shown in Table 4.

[0131] Table 4

[0132] Table 4 shows that solution a of Example 1 is more suitable for use in facial cleanser than other rhamnolipid raw materials.

[0133] Shampoo

[0134] The three different di-rhamnolipid ratio daily-use chemical raw materials (solutions a / b / c) obtained in Example 1 and the rhamnolipid crude product obtained in the comparative example were formulated into shampoos according to the formulation of Table 5.

[0135] Table 5

[0136] Processing procedure:

[0137] The above raw materials were added in the formulation ratio, stirred at 35°C for 30 minutes, then the solution pH was adjusted to 6.5 with citric acid, and cooled to room temperature.

[0138] Ten consumers were selected to score the cleansing power, smoothness, oil control, and gloss of the shampoos respectively added with solutions a / b / c of Example 1 and the rhamnolipid crude product of the comparative example, with a full score of 10. The results are shown in Table 6.

[0139] Table 6

[0140] Table 6 shows that solution b of Example 1 is more suitable for use in shampoo than other rhamnolipid raw materials.

[0141] Laundry liquid

[0142] The three different di-rhamnolipid ratio daily-use chemical raw materials (solutions a / b / c) obtained in Example 1 and the rhamnolipid crude product obtained in the comparative example were formulated into laundry liquids according to the formulation of Table 7.

[0143] Table 7

[0144] Processing procedure:

[0145] The above raw materials were added in the formulation ratio, stirred at 35°C for 30 minutes, then the solution pH was adjusted to 7.8 with sodium hydroxide, and cooled to room temperature.

[0146] The stain removal, fabric softness, and color protection of the laundry liquids respectively added with solutions a / b / c of Example 1 and the rhamnolipid crude product of the comparative example were scored, with a full score of 10. The results are shown in Table 8.

[0147] Table 8

[0148] Table 8 shows that solution c of Example 1 is more suitable for laundry liquid compared to other rhamnolipid feedstocks.

[0149] The above experiment was performed using the three different di-rhamnolipid ratios of the personal care feedstocks obtained in Example 2 (solutions a / b / c) and the same results were obtained, i.e. solution a is more suitable for facial wash, solution b is more suitable for shampoo and solution c is more suitable for laundry liquid.

[0150] Obviously, the above examples are merely illustrative in nature and are not intended to limit the scope of the application. Various changes or modifications can be suggested to one skilled in the art, and it is intended that the application encompass such changes and modifications as fall within the scope of the appended claims.

Claims

1. A process for producing rhamnolipid cosmetic raw material, comprising the following steps: (1) culturing Pseudomonas aeruginosa in fermentation, controlling the pH in the fermentation process at 7.0-8.0, to obtain a fermentation liquor with a rhamnolipid content of 60-80 g / L and a double glycolipid proportion in the rhamnolipid of 60-70%; (2) removing the bacteria in the fermentation liquor using a solid-liquid separation device to obtain a sterilized fermentation liquor; (3) adding acid to the sterilized fermentation liquor to adjust the pH to 3.0-3.5, and separating the solid and liquid phases to obtain a solid A and a supernatant A; further adding acid to the supernatant A to reduce the pH by 0.5, and separating the solid and liquid phases to obtain a solid B and a supernatant B; further adding acid to the supernatant B to reduce the pH by 0.5 again, and separating the solid and liquid phases to obtain a solid C and a supernatant C.

2. The production process according to claim 1, characterized in that: The solid A has a rhamnolipid mass percentage of 80-93%, and a double glycolipid mass percentage in the rhamnolipid of 90-95%; The solid B has a rhamnolipid mass percentage of 80-93%, and a double glycolipid mass percentage in the rhamnolipid of 70-80%; and / or The solid C has a rhamnolipid mass percentage of 80-93%, and a double glycolipid mass percentage in the rhamnolipid of 35-50%.

3. The production process according to claim 1 or 2, characterized in that: In step (1), the fermentation culture uses soybean oil as the carbon source and yeast powder as the nitrogen source, and the fermentation temperature is 30-37°C; and / or In step (3), the acid is an organic acid and / or an inorganic acid; The organic acid is preferably one or more of citric acid, formic acid, acetic acid, valeric acid, and butyric acid, and is more preferably acetic acid; The inorganic acid is preferably one or more of sulfuric acid, hydrochloric acid, nitric acid, and phosphoric acid, and is more preferably sulfuric acid.

4. The production process according to any one of claims 1 to 3, characterized in that: Further comprising: (4) dissolving the solid A, solid B, and solid C in an alcohol solvent respectively, filtering to obtain a filtrate, and evaporating the solvent in the filtrate to obtain a solid a, solid b, and solid c, respectively.

5. The production process according to any one of claims 1 to 4, characterized in that: Further comprising: (5) adding to the solid a an aqueous KOH solution with a mass percentage of 5-6% and / or an aqueous NaOH solution with a mass percentage of 5-6% in an amount of 0.9-1.1 times the mass of the solid a, and 1,3-butanediol, glycerol, and / or 1,2-pentanediol in an amount of 0.5-0.55 times the mass of the solid a, to obtain a solution a with a pH of 5.5-6.0, a rhamnolipid mass percentage of 35-40%, and a double glycolipid mass percentage in the rhamnolipid of 90-95%; and / or (6) adding to the solid b an aqueous KOH solution with a mass percentage of 5-6% and / or an aqueous NaOH solution with a mass percentage of 5-6% in an amount of 1.2-1.25 times the mass of the solid b, and 1,3-butanediol, glycerol, and / or 1,2-pentanediol in an amount of 0.6-0.65 times the mass of the solid b, to obtain a solution b with a pH of 6.0-6.5, a rhamnolipid mass percentage of 32-35%, and a double glycolipid mass percentage in the rhamnolipid of 70-80%; and / or In the solid c, 1.3-1.4 times of the mass of the solid c is added with 5-6% mass percentage KOH aqueous solution and / or 5-6% mass percentage NaOH aqueous solution and 0.7-0.75 times of the mass of the solid c is added with 1,3-butanediol, glycerol and / or 1,2-pentanediol, to obtain a solution c with pH 6.5-7.0, the mass percentage of rhamnolipid in the solution c is 30-33%, and the mass percentage of di-rhamnolipid in the rhamnolipid is 35-50%.

6. The production process according to claim 5, characterized in that: In the step (5), the pH value of the solution a is resistant to 5-6, and the CMC is 0.005-0.007 mol / L; The pH value of the solution b is resistant to 6.5-7.5, and the CMC is 0.008-0.01 mol / L; and / or The pH value of the solution c is resistant to 7.5-8.5, and the CMC is 0.012-0.02 mol / L.

7. The solid A, solid B, solid C, solid a, solid b, solid c, solution a, solution b and solution c obtained by the production process of any one of claims 1-6.

8. The use of the solid A, solid a and / or solution a of claim 7 as a daily chemical raw material in the preparation of skin care products in personal care products.

9. The use of the solid B, solid b and / or solution b of claim 7 as a daily chemical raw material in the preparation of hair care products in personal care products.

10. The use of the solid C, solid c and / or solution c of claim 7 as a daily chemical raw material in the preparation of household care products.

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